title: Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays.
abstract: The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TelN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TelN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes.
authors.affiliation: School of Science, Monash University, Bandar Sunway, Malaysia.
authors.forename: Qingwen
authors.initials: Q
authors.lastname: Chen
authors.forename: Kumaran
authors.initials: K
authors.lastname: Narayanan
mesh_terms: D001419:Bacteria / Q000821:virology; D001435:Bacteriophages / Q000201:enzymology*; D001671:Biochemistry / Q000379:methods*; D004926:Escherichia coli / Q000821:virology; D019098:Telomerase / Q000302:isolation & purification* / Q000378:metabolism; D016615:Telomere / Q000737:chemistry
chemical_list: D019098:Telomerase